5 Tips for Setting Up Your PCR

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1.   Choose a polymerase that matches your need

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2.     Take time to carefully to design your primers

 

     Primer design

l  20-30 nucleotides

l  GC content 40-60%

l  Tms witnin with 5 degrees of each other

 

     3.Calculate annealing temperature

      *Always recalculate annealing temperature when switching enzymes

 

     4.Calculate GC content of your target

     5.Setup on ice and preheat thermoclcler

      Medicalet Centrifuge tube with

l  Room temperature reaction setup

l  Increased specificity